The present invention relates to acid derivatives that are inhibitors of serine proteases such as Factor VIIa, Factor IXa, Factor Xa, Factor FXIa, tryptase, and urokinase. These acid derivatives are useful as anticoagulants in treating and preventing cardiovascular diseases, as anti-inflammatory agents, and as metastasis inhibitors in treating cancer.
Under normal conditions, the coagulation system is naturally balanced in favor of anticoagulation by a number of proteins circulating in the blood. These proteins include antithrombin III, a serine-protease inhibitor, and protein C, a vitamin-K dependent protein formed in the liver. When injury or trauma occurs, thrombin is produced at precise levels through an ordered series of reactions. Thrombin is a proteolytic enzyme that occupies a central position in the coagulation process. Thrombin catalyzes the conversion of fibrinogen to fibrin, is a key effector enzyme for blood clotting, and also is pivotal for other functions, such as activation of helper proteins (including Factors V and VIII and thrombomodulin), and its own activation. Disturbances in the natural balance between pro- and anti-coagulant forces may result in bleeding or thrombotic diseases.
The series of reactions leading to thrombin production involve a number of coagulation factors present in the blood as precursors (e.g., Factors VII-XII). When the coagulation system is triggered (e.g., when trauma occurs), the coagulation factors are transformed into activated factors (e.g., Factors VIIa, IXa, Xa, XIa, etc.). Factor VII forms a complex with a membrane protein called tissue factor, to which Factor VIIa tightly binds. Thus, Factor VIIa is present as a complex bound to tissue factor. When triggered, the coagulation factors and tissue factor complexes undergo an ordered chain of reactions that ultimately lead to conversion of Factor X to Factor Xa, and Factor Xa catalyzes the conversion of prothrombin to thrombin.
An elevated plasma level of coagulation factors, particularly Factor VIIa, is a risk factor for fatal myocardial infarction and associated with coronary artery disease and other abnormalities of the coagulation system, e.g., thrombosis, ischemic vascular disease, intravascular clotting, stroke, embolisms, and so forth. Accordingly, antithrombotic agents have been researched and developed for use in treating cardiovascular and other diseases. Presently established antithrombotic agents include heparin, coumarin, and aspirin, among others. There are, however, limitations with these agents. For example, both heparin and coumarin have a highly-variable dose-related response, and their anticoagulant effects must be closely monitored to avoid a risk of serious bleeding. The erratic anticoagulant response of heparin is likely due to its propensity to bind non-specifically to plasma proteins. Aspirin has a limited efficacy and at high doses presents a risk of gastrointestinal bleeding. Thrombin inhibitors and their drawbacks are further discussed in WO 96/20689 to duPont Merck Pharmaceutical Co.
As may be appreciated, those in the field of pharmaceutical research continue to seek to develop new compounds and compositions having increased effectiveness and bioavailability and/or having fewer side effects. See, e.g., Jakobsen et al., xe2x80x9cInhibitors of the Tissue Factor/Factor VIIa-induced Coagulation: Synthesis and In vitro Evaluation of Novel Specific 2-aryl Substituted 4H-3,1-benzoxazin-4-ones,xe2x80x9d Bioorganic and Medicinal Chemistry, Vol. 8 (August 2000), at pp. 2095-2103; and J. Hirsh et al., xe2x80x9cThrombosis, New Antithrombotic Agents,xe2x80x9d Lancet, Vol. 353 (Apr. 24, 1999), at pp. 1431-36. There is particularly an interest in developing agents that can selectively and directly inhibit key factors in the complicated coagulation process. Compounds effective in inhibiting Factor Xa are described in U.S. Pat. application Ser. No. 09/478,632, filed Jan. 6, 2000, Ser. No. 09/633,751, filed Aug. 7, 2000, and Ser. No. 09/496,571, filed Feb. 2, 2000. Compounds effective in inhibiting Factors VIIa, Xa, as well as tryptase and urokinase are described in U.S. patent application Ser. No. 09/458,847, filed Dec. 13, 1999. The above referenced ""632, ""751, ""571, and ""847 applications show lactam compounds and are each assigned to the present assignee with common inventors herewith. Factor Xa inhibitors are also disclosed in PCT applic. WO 98/57937 to the duPont Merck Pharmaceutical Co.
PCT patent application WO 99/41231 to Ono Pharmaceuticals Inc., (xe2x80x9cOnoxe2x80x9d) discloses a series of amidino derivatives such as 2-(3-(4-amidinophenylcarbamoyl)-naphthalen-2-yl)-5-((2,2-methylpropyl)carbamoyl benzoic acid, which are claimed to be Factor VIIa inhibitors. The Ono application is discussed in Kohrt et al., xe2x80x9cAn Efficient Synthesis of 2-(3-(4-Amidinophenylcarbamoyl)naphthalen-2-yl)-5-((2,2-methylpropyl)carbamoyl benzoic acid: a Factor VIIa Inhibitor Discovered by the Ono Pharmaceutical Company,xe2x80x9d Tetrahedron Letters, Vol. 41 (June 2000), at pp. 6041-44, which reports that Ono fails to fully describe an effective method for making the titled compound. Inhibitors of Factor VIIa are also reported in WO 01/44172 to Axys Pharm. Inc. PCT patent application WO 98/47876 to Akzo Novel N. V., published Oct. 29, 1998, discloses certain bicyclic groups such as isoquinoline groups which reportedly are advantageous for promoting pharmacological properties, and isoquinoline-containing compounds are disclosed in WO 94/29273 to SmithKline Beecham Corp. Biphenyl compounds and/or acid substituted bicyclic compounds are also disclosed in U.S. Pat. Nos. 5,612,341, 6,248,767 B1, 3,995,045, EP patent application 0 206 567 A2 to Warner Lambert Co., and WO 01/70678 to Merck Patent GmbH.
The patents, patent applications, and articles cited above are incorporated herein by reference.
The present invention provides acid-based compounds useful as inhibitors of Factor VIIa, Factor IXa, Factor Xa, Factor FXIa, tryptase, and urokinase.
Acid derivatives are provided that are inhibitors of serine proteases having the Formula I: 
or pharmaceutically-acceptable salts, hydrates or prodrugs thereof, wherein:
W is selected from C2-10alkyl, C2-10alkenyl, substituted C2-10alkyl, substituted C2-10alkenyl, xe2x80x94C(xe2x95x90O)NR4R5, xe2x80x94OR6, xe2x80x94CO2R4, xe2x80x94C(xe2x95x90O)R4, xe2x80x94SR4, xe2x80x94S(O)pR4, xe2x80x94NR4R5, xe2x80x94NR4SO2R5, xe2x80x94NR4aSO2NR4R5, xe2x80x94NR4CO2R5, xe2x80x94NR4C(xe2x95x90O)R5, xe2x80x94NR4aC(xe2x95x90O)NR4R5, xe2x80x94SO2NR4R5, heterocyclo, heteroaryl, aryl, and cycloalkyl;
ring B is phenyl or pyridyl;
X2 is N, CH, or C, provided that X2 is C when R1 and R2 join to form a fully unsaturated ring;
L is xe2x80x94(CR18R19)sxe2x80x94Yxe2x80x94(CR18aR19a)t;
Y is selected from xe2x80x94C(xe2x95x90O), xe2x80x94C(xe2x95x90O)NR13xe2x80x94, xe2x80x94NR13C(xe2x95x90O)xe2x80x94, xe2x80x94NR13CR14R15xe2x80x94, xe2x80x94CR14R15xe2x80x94NR13xe2x80x94, and xe2x80x94CR13R14xe2x80x94CR15R16xe2x80x94;
Z is a 5 to 7-membered monocyclic or 8 to 11-membered bicyclic aryl, heteroaryl, heterocyclo, or cycloalkyl, wherein each Z group is optionally substituted with up to two R20 and/or up to one R21, except Z is not phenyl substituted with phenyloxy when W is methoxy, s is 0 and Y is xe2x80x94CH2xe2x80x94CH2xe2x80x94;
R1 and R2 (i) are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, heteroaryl, aryl, heterocyclo, and cycloalkyl; or (ii) are taken together to form an aryl, heteroaryl, cycloalkyl, or heterocyclo, provided that R1 and R2 do not together form pyrazole when W is methoxy and Z is biphenyl; and when R1 and R2 individually or together form a heteroaryl, aryl, heterocyclo, or cycloalkyl, said cyclic group is optionally substituted with up to three R26;
R3 is hydrogen, alkyl, substituted alkyl, heteroaryl, aryl, heterocyclo, cycloalkyl, or alkyl substituted with xe2x80x94OC(xe2x95x90O)R24 or xe2x80x94OC(xe2x95x90O)OR24, wherein R24 is alkyl, substituted alkyl, or cycloalkyl, provided that R3 is not phenyl when W is methoxy;
R4, R4a, R5 and R6 are (i) independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, heteroalkyl, substituted heteroalkyl, aryl, heteroaryl, heterocyclo, and cycloalkyl; or alternatively, (ii) R4 and R5 may be taken together to form a five-to-seven membered heteroaryl or heterocyclo, except when W is xe2x80x94S(O)pR4, then R4 is not hydrogen;
R8 and R26 (i) are at each occurrence independently selected from hydrogen, OR30, NR31R32, NR31SO2R32a, alkyl, alkenyl, substituted alkyl, substituted alkenyl, halogen, haloalkyl, haloalkoxy, cyano, nitro, alkylthio, xe2x80x94C(xe2x95x90O)H, acyl, xe2x80x94CO2H, alkoxycarbonyl, sulfonamido, sulfonyl, and phenyl, or (ii) two of R8 and/or two of R26 may be taken together to form a fused benzo ring, a fused heteroaryl, a fused cycloalkyl, or a fused heterocyclo other than a five or six membered heterocyclo having as its heteroatoms two oxygen atoms, provided further that when two R26 form a fused benzo ring, then Z is not phenyl substituted in the para position with cyano or a five-membered heterocycle or heteroaryl;
R13, R14, R15, R16, R18, R18a, R19, and R19a are selected from hydrogen, lower alkyl, hydroxy, and lower alkyl substituted with hydroxy or halogen;
R20 and R21 are independently selected at each occurrence from hydrogen, halogen, alkyl, substituted alkyl, haloalkyl, haloalkoxy, cyano, nitro, xe2x80x94C(xe2x95x90O)NR22R23, xe2x80x94OR22, xe2x80x94CO2R22, xe2x80x94C(xe2x95x90O)R22, xe2x80x94SR22, xe2x80x94S(O)qR22a, xe2x80x94NR22R23, xe2x80x94NR22SO2R23 NR22, xe2x80x94NR22CO2R23, xe2x80x94NR22C(xe2x95x90O)R23, xe2x80x94NR22C(xe2x95x90O)NR23R33, xe2x80x94SO2NR22R23, xe2x80x94NR22SO2NR23R33, five or six membered heterocyclo or heteroaryl, phenyl, and four to seven membered cycloalkyl, wherein when R20 and/or R21 independent of each other comprise a cyclic group, each cyclic group in turn is optionally substituted with up to three of C1-4alkyl, C1-4alkoxy, halogen, hydroxy, haloalkyl, haloalkoxy, amino, alkylamino, and/or cyano;
R22, R23 and R33 are independently selected from hydrogen, alkyl, and substituted alkyl;
R22a is alkyl or substituted alkyl;
R30 at each occurrence is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, and phenyl;
R31 and R32 at each occurrence are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, and cycloalkyl;
R32a is alkyl, substituted alkyl, alkenyl, substituted alkenyl, or cycloalkyl;
m is 0, 1 or 2 when ring B is phenyl and 0 or 1 when ring B is pyridyl;
p and q are independently 1 or 2; and
s and t are independently 0, 1 or 2.
The compounds of this invention are surprisingly selective inhibitors of serine proteases. For example, it has been found that certain selections for the groups xe2x80x9cZxe2x80x94Lxe2x80x94xe2x80x9d in formula I, provide compounds which are particularly selective for inhibition of one or more serine proteases versus other proteases. To illustrate, it has been surprisingly found that when Zxe2x80x94Lxe2x80x94 is selected from: 
compounds of formula I are particularly selective for inhibition of FVIIa.
As another illustration, it has been found that when Zxe2x80x94Lxe2x80x94 is 
compounds of formula I are particularly selective for inhibition of FXa.
Included within the scope of the invention are pharmaceutical compositions for treating a serine protease disease, an inflammatory or immune condition, or cancer, comprising at least one compound of formula I or a pharmaceutically acceptable salt, hydrate or prodrug thereof, and a pharmaceutically acceptable carrier or diluent. Also included in the invention are methods of treating such diseases comprising administering to a mammal in need of such treatment at least one compound of formula I or a pharmaceutically acceptable salt, hydrate or prodrug thereof. Further included in the invention are compositions for use as anticoagulants during the preparation, use, storage, or fractionation of blood and methods of maintaining blood in the fluid phase during its preparation, use, storage, or fractionation.
The following are definitions of terms used in this specification. The initial definition provided for a group or term herein applies to that group or term throughout this specification, individually or as part of another group, unless otherwise indicated.
The term xe2x80x9calkylxe2x80x9d refers to straight or branched chain hydrocarbon groups having 1 to 12 carbon atoms, preferably 1 to 8 carbon atoms. Lower alkyl groups, that is, alkyl groups of 1 to 4 carbon atoms, are most preferred.
When numbers appear in a subscript after the symbol xe2x80x9cCxe2x80x9d, the subscript defines with more specificity the number of carbon atoms that a particular group may contain. For example, xe2x80x9cC1-6alkylxe2x80x9d refers to straight and branched chain alkyl groups with one to six carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, n-pentyl, and so forth.
The term xe2x80x9csubstituted alkylxe2x80x9d refers to an alkyl group as defined above having one, two, or three substituents selected from the group consisting of halo, alkenyl, alkynyl, nitro, cyano, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, xe2x80x94CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, xe2x80x94S(O)2(alkyl), keto (xe2x95x90O), aryl, heteroaryl, heterocyclo, and cycloalkyl, including phenyl, benzyl, phenylethyl, phenyloxy, and phenylthio. The substituents for xe2x80x9csubstituted alkylxe2x80x9d groups may also be selected from the group consisting of xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, and xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, wherein each of Rxe2x80x2 and Rxe2x80x3 is independently selected from hydrogen, alkyl, cycloalkyl, and alkyl substituted with one to two of alkenyl, halogen, haloalkyl, haloalkoxy, cyano, nitro, hydroxy, alkoxy, alkylthio, amino, alkylamino, phenyl, benzyl, phenyloxy, and benzyloxy. Alternatively, Rxe2x80x2 and Rxe2x80x3 may together form a heterocyclo or heteroaryl ring. When a substituted alkyl includes an aryl, heterocyclo, cycloalkyl, or heteroaryl substituent, said ringed systems are as defined below and thus may have zero, one, two, or three substituents, also as defined below.
When the term xe2x80x9calkylxe2x80x9d is used in conjunction with another group, e.g., arylalkyl, hydroxyalkyl, etc., the term defines with more specificity a particular substituent that a substituted alkyl will contain. For example, arylalkyl refers to a substituted alkyl group having from 1 to 12 carbon atoms and at least one aryl substituent, and xe2x80x9clower arylalkylxe2x80x9d refers to substituted alkyl groups having 1 to 4 carbon atoms and at least one aryl substituent.
The term xe2x80x9calkenylxe2x80x9d refers to straight or branched chain hydrocarbon groups having 2 to 12 carbon atoms and at least one double bond. Alkenyl groups of 2 to 6 carbon atoms and having one double bond are most preferred.
The term xe2x80x9calkynylxe2x80x9d refers to straight or branched chain hydrocarbon groups having 2 to 12 carbon atoms and at least one triple bond. Alkynyl groups of 2 to 6 carbon atoms and having one triple bond are most preferred.
The term xe2x80x9calkylenexe2x80x9d refers to bivalent straight or branched chain hydrocarbon groups having 1 to 12 carbon atoms, preferably 1 to 8 carbon atoms, e.g., {xe2x80x94CH2xe2x80x94}n, wherein n is 1 to 12, preferably 1-8. Lower alkylene groups, that is, alkylene groups of 1 to 4 carbon atoms, are most preferred. The terms xe2x80x9calkenylenexe2x80x9d and xe2x80x9calkynylenexe2x80x9d refer to bivalent radicals of alkenyl and alknyl groups, respectively, as defined above.
When reference is made to a substituted alkylene, alkenylene, or alkynylene group, these groups are substituted with one to three substitutents as defined above for alkyl groups. A ringed substituent of an alkyl, alkenyl, alkynyl, alkylene, alkenylene, or alkynylene may be joined at a terminal atom or an available intermediate (branch or chain) atom and thus may comprise, for example, the groups 
and so forth.
The term xe2x80x9calkoxyxe2x80x9d refers to an alkyl group as defined above having one, two or three oxygen atoms (xe2x80x94Oxe2x80x94) in the alkyl chain. For example, the term xe2x80x9calkoxyxe2x80x9d includes the groups xe2x80x94Oxe2x80x94C1-12alkyl, xe2x80x94C1-6alkylene-Oxe2x80x94C1-6alkyl, xe2x80x94C1-4alkylene-Oxe2x80x94C1-4alkylene-Oxe2x80x94C1-4alkyl, Oxe2x80x94C1-4alkylene-Oxe2x80x94C1-4alkylene-Oxe2x80x94C1-4alkyl, and so forth.
The term xe2x80x9calkylthioxe2x80x9d refers to an alkyl group as defined above bonded through one or more sulfur (xe2x80x94Sxe2x80x94) atoms. For example, the term xe2x80x9calkylthioxe2x80x9d includes the groups xe2x80x94Sxe2x80x94C1-12alkyl, xe2x80x94S1-6alkylene-Sxe2x80x94C1-6alkyl, etc.
The term xe2x80x9calkylaminoxe2x80x9d refers to an alkyl group as defined above bonded through one or more nitrogen (xe2x80x94NRxe2x80x94) groups. The term alkylamino refers to straight and branched chain groups and thus, for example, includes the groups xe2x80x94NH(C1-12alkyl) and xe2x80x94N(C1-6alkyl)2. 
When a subscript is used with reference to an alkoxy, alkylthio or alkylamino, the subscript refers to the number of carbon atoms in the group in addition to heteroatoms. Thus, for example, monovalent C1-2alkylamino includes the groups xe2x80x94NHxe2x80x94CH3, xe2x80x94NHxe2x80x94CH2xe2x80x94CH3, and xe2x80x94Nxe2x80x94(CH3)2. A lower alkylamino comprises an alkylamino having from one to four carbon atoms.
When reference is made to a substituted alkoxy or alkylthio, the carbon atoms of said groups are substituted with one to three substituents as defined above for alkyl groups. When reference is made to a substituted alkylamino, the carbon and/or nitrogen atoms of these groups are substituted with one to three substitutents appropriately selected from the group of substituents recited above for alkyl groups. Additionally, the alkoxy, alkylthio, or alkylamino groups may be monovalent or bivalent. By xe2x80x9cmonovalentxe2x80x9d it is meant that the group has a valency (i.e., power to combine with another group), of one, and by xe2x80x9cbivalentxe2x80x9d it is meant that the group has a valency of two. Thus, for example, a monovalent alkoxy includes groups such as xe2x80x94Oxe2x80x94C1-12alkyl and xe2x80x94C1-6alkylene-Oxe2x80x94C1-6alkyl, whereas a bivalent alkoxy includes groups such as xe2x80x94Oxe2x80x94C1-12alkylene- and xe2x80x94C1-6alkylene-Oxe2x80x94C1-6alkylene-, etc.
The term xe2x80x9cheteroalkylxe2x80x9d is used herein to refer saturated and unsaturated straight or branched chain hydrocarbon groups having 2 to 12 carbon atoms, preferably 2 to 8 carbon atoms, wherein one, two or three carbon atoms in the straight chain are replaced by a heteroatom (O, S or N). Thus, the term xe2x80x9cheteroalkylxe2x80x9d includes alkoxy, alkylthio, and alkylamino groups, as defined above, as well as alkyl groups having a combination of heteroatoms selected from O, S, or N. A xe2x80x9cheteroalkylxe2x80x9d herein may be monovalent or bivalent, and for example, may comprise the groups xe2x80x94Oxe2x80x94(CH2)2-5NHxe2x80x94(CH2)2xe2x80x94 or xe2x80x94Oxe2x80x94(CH2)2-5NHxe2x80x94CH3, etc. A xe2x80x9csubstituted heteroalkylxe2x80x9d has to three substituents appropriately selected from those recited above for alkyl groups.
The term xe2x80x9cacylxe2x80x9d refers to a carbonyl group 
linked to an organic radical including an alkyl, alkenyl, alkynyl, substituted alkyl, substituted alkenyl, or substituted alkynyl group, as defined above.
The term xe2x80x9calkoxycarbonylxe2x80x9d refers to a carboxy or ester group 
linked to an organic radical including an alkyl, alkenyl, alkynyl, substituted alkyl, substituted alkenyl, or substituted alkynyl group, as defined above.
The term xe2x80x9chaloxe2x80x9d or xe2x80x9chalogenxe2x80x9d refers to chloro, bromo, fluoro and iodo.
The term xe2x80x9chaloalkylxe2x80x9d means an alkyl having one or more halo substituents, e.g., including trifluoromethyl.
The term xe2x80x9chaloalkoxyxe2x80x9d means an alkoxy group having one or more halo substituents. For example, xe2x80x9chaloalkoxyxe2x80x9d includes xe2x80x94OCF3.
The term xe2x80x9csulfonylxe2x80x9d refers to a sulphoxide group (i.e., xe2x80x94S(O)1-2xe2x80x94) linked to an organic radical including an alkyl, alkenyl, alkynyl, substituted alkyl, substituted alkenyl, or substituted alkynyl group, as defined above. The organic radical to which the sulphoxide group is attached may be monovalent (e.g., xe2x80x94SO2-alkyl), or bivalent (e.g., xe2x80x94SO2-alkylene, etc.)
The term xe2x80x9csulfonamidexe2x80x9d refers to the group xe2x80x94S(O)2NRxe2x80x2Rxe2x80x3, wherein Rxe2x80x2 and Rxe2x80x3 may be hydrogen or alkyl, alkenyl, alkynyl, substituted alkyl, substituted alkenyl, or substituted alkynyl, as defined above. Rxe2x80x2 and Rxe2x80x3 may be monovalent or bivalent (e.g., xe2x80x94SO2xe2x80x94NH-alkylene, etc.)
The term xe2x80x9carylxe2x80x9d refers to phenyl, biphenyl, 1-naphthyl and 2-naphthyl, with phenyl being preferred. The term xe2x80x9carylxe2x80x9d includes such rings having zero, one, two or three substituents selected from the group consisting of halo, alkyl, alkenyl, alkynyl, nitro, cyano, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, phenyl, benzyl, phenylethyl, phenyloxy, phenylthio, cycloalkyl, heterocyclo, heteroaryl, xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, and/or alkyl substituted with one to three of halo, nitro, cyano, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, phenyl, benzyl, phenylethyl, phenyloxy, phenylthio, cycloalkyl, heterocyclo, heteroaryl, xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, and/or xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, wherein each of Rxe2x80x2 and Rxe2x80x3 is independently selected from hydrogen, alkyl, alkoxy, hydroxyalkyl, and arylalkyl, or Rxe2x80x2 and Rxe2x80x3 together form a heterocyclo or heteroaryl ring. When an aryl is substituted with a further ring, said ring may in turn be substituted with one to three of halogen, haloalkyl, haloalkoxy, cyano, nitro, hydroxy, alkoxy, alkylthio, amino, alkylamino, phenyl, benzyl, phenyloxy, and benzyloxy.
The term xe2x80x9ccycloalkylxe2x80x9d refers to fully saturated and partially unsaturated hydrocarbon rings of 3 to 9, preferably 3 to 7 carbon atoms. The term xe2x80x9ccycloalkylxe2x80x9d includes such rings having zero, one, two, or three substituents, preferably zero or one, selected from the group consisting of halo, alkyl, alkenyl, alkynyl, nitro, cyano, oxo (xe2x95x90O), hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, keto, xe2x95x90Nxe2x80x94OH, xe2x95x90Nxe2x80x94O-alkyl, heteroaryl, heterocyclo, a five or six membered ketal (i.e. 1,3-dioxolane or 1,3-dioxane), a four to seven membered carbocyclic ring, xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, and/or alkyl substituted with one to three of halo, nitro, cyano, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, phenyl, benzyl, phenylethyl, phenyloxy, phenylthio, a four to seven membered carbocyclic ring, heterocyclo, heteroaryl, xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, and/or xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, wherein each of Rxe2x80x2 and Rxe2x80x3 is independently selected from hydrogen, alkyl, alkoxy, hydroxyalkyl, and arylalkyl, or Rxe2x80x2 and Rxe2x80x3 together form a heterocyclo or heteroaryl ring. When a cycloalkyl is substituted with a further ring, said ring may in turn be substituted with one to three of halogen, haloalkyl, haloalkoxy, cyano, nitro, hydroxy, alkoxy, alkylthio, amino, alkylamino, phenyl, benzyl, phenyloxy, and benzyloxy.
The term xe2x80x9cheterocycloxe2x80x9d refers to substituted and unsubstituted non-aromatic 3 to 7 membered monocyclic groups, 7 to 11 membered bicyclic groups, and 10 to 15 membered tricyclic groups which have at least one heteroatom (O, S or N) in at least one of the rings. Each ring of the heterocyclo group containing a heteroatom can contain one or two oxygen or sulfur atoms and/or from one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less, and further provided that the ring contains at least one carbon atom. The fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or unsaturated. The nitrogen and sulfur atoms may optionally be oxidized and the nitrogen atoms may optionally be quaternized. The heterocyclo group may be attached at any available nitrogen or carbon atom. The heterocyclo ring may contain zero, one, two or three substituents selected from the group consisting of halo, alkyl, alkenyl, alkynyl, nitro, cyano, oxo, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, keto, xe2x95x90Nxe2x80x94OH, xe2x95x90Nxe2x80x94O-alkyl, aryl, heteroaryl, cycloalkyl, a five or six membered ketal (i.e. 1,3-dioxolane or 1,3-dioxane), xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, and/or alkyl substituted with one to three of halo, nitro, cyano, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, phenyl, benzyl, phenylethyl, phenyloxy, phenylthio, cycloalkyl, heterocyclo, heteroaryl, xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, and/or xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, wherein each of Rxe2x80x2 and Rxe2x80x3 is independently selected from hydrogen, alkyl, alkoxy, hydroxyalkyl, and arylalkyl, or Rxe2x80x2 and Rxe2x80x3 together form a heterocyclo or heteroaryl ring. When a heterocyclo is substituted with a further ring, said ring may in turn be substituted with one to three of halogen, haloalkyl, haloalkoxy, cyano, nitro, hydroxy, alkoxy, alkylthio, amino, alkylamino, phenyl, benzyl, phenyloxy, and benzyloxy.
Exemplary monocyclic groups include azetidinyl, pyrrolidinyl, oxetanyl, imidazolinyl, oxazolidinyl, isoxazolinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl and the like. Exemplary bicyclic heterocyclo groups include quinuclidinyl.
The term xe2x80x9cheteroarylxe2x80x9d refers to substituted and unsubstituted aromatic 5 or 6 membered monocyclic groups, 9 or 10 membered bicyclic groups, and 11 to 14 membered tricyclic groups which have at least one heteroatom (O, S or N) in at least one of the rings. Each ring of the heteroaryl group containing a heteroatom can contain one or two oxygen or sulfur atoms and/or from one to four nitrogen atoms provided that the total number of heteroatoms in each ring is four or less and each ring has at least one carbon atom. The fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or unsaturated. The nitrogen and sulfur atoms may optionally be oxidized and the nitrogen atoms may optionally be quaternized. Heteroaryl groups which are bicyclic or tricyclic must include at least one fully aromatic ring but the other fused ring or rings may be aromatic or non-aromatic. The heteroaryl group may be attached at any available nitrogen or carbon atom of any ring. The heteroaryl ring system may contain zero, one, two or three substituents selected from the group consisting of halo, alkyl, alkenyl, alkynyl, nitro, cyano, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, phenyl, benzyl, phenylethyl, phenyloxy, phenylthio, cycloalkyl, heterocyclo, a further monocyclic heteroaryl, xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, and/or alkyl substituted with one to three of halo, nitro, cyano, hydroxy, alkoxy, alkylthio, xe2x80x94CO2H, xe2x80x94C(xe2x95x90O)H, CO2-alkyl, xe2x80x94C(xe2x95x90O)alkyl, phenyl, benzyl, phenylethyl, phenyloxy, phenylthio, cycloalkyl, heterocyclo, heteroaryl, xe2x80x94NRxe2x80x2Rxe2x80x3, xe2x80x94C(xe2x95x90O)NRxe2x80x2Rxe2x80x3, xe2x80x94CO2NRxe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2CO2xe2x80x2Rxe2x80x3, xe2x80x94NRxe2x80x2C(xe2x95x90O)Rxe2x80x3, xe2x80x94SO2NRxe2x80x2Rxe2x80x3, and/or xe2x80x94NRxe2x80x2SO2xe2x80x2Rxe2x80x3, wherein each of Rxe2x80x2 and Rxe2x80x3 is independently selected from hydrogen, alkyl, alkoxy, hydroxyalkyl, and arylalkyl, or Rxe2x80x2 and Rxe2x80x3 together form a heterocyclo or heteroaryl ring. When a heteroaryl is substituted with a further ring, said ring may in turn be substituted with one to three of halogen, haloalkyl, haloalkoxy, cyano, nitro, hydroxy, alkoxy, alkylthio, amino, alkylamino, phenyl, benzyl, phenyloxy, and benzyloxy.
Exemplary monocyclic heteroaryl groups include pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl and the like.
Exemplary bicyclic heteroaryl groups include indolyl, benzothiazolyl, benzodioxolyl, benzoxaxolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridinyl, dihydroisoindolyl, tetrahydroquinolinyl and the like.
Exemplary tricyclic heteroaryl groups include carbazolyl, benzidolyl, phenanthrollinyl, acridinyl, phenanthridinyl, xanthenyl and the like.
The term xe2x80x9ccarbocyclicxe2x80x9d refers to optionally substituted aromatic or non-aromatic 3 to 7 membered monocyclic and 7 to 11 membered bicyclic groups, in which all atoms of the ring or rings are carbon atoms.
When the term xe2x80x9cunsaturatedxe2x80x9d is used herein to refer to a ring or group, the ring or group may be fully unsaturated or partially unsaturated.
The term xe2x80x9cmetal ionxe2x80x9d refers to alkali metal ions such as sodium, potassium or lithium and alkaline earth metal ions such as magnesium and calcium, as well as zinc and aluminum.
Whenever a bond appears in a formula as a dashed-double bond, i.e., with one bond appearing as a dash as in 
it should be understood that such bonds may be selected from single or double bonds, as appropriate given the selections for adjacent atoms and bonds. For example, in formula I, above, when X2 is N or CH, the bonds linking R1 to X2 and X2 to C6 are single bonds; and when X2 is C, one of the bonds linking X2 to an adjacent atom is a double bond, i.e., either a bond to R1 or to C6 is a double bond.
It should be understood that one skilled in the field may make various substitutions for each of the groups recited in the claims herein, without departing from the spirit or scope of the invention. For example, one skilled in the field may replace a W group recited in the claims with a cyano, halogen, or methyl group. The linker group xe2x80x9cLxe2x80x9d recited in the claims may be replaced with the group xe2x80x94(Rxe2x80x2)uxe2x80x94Yxe2x80x2xe2x80x94(Rxe2x80x3)vxe2x80x94 wherein Yxe2x80x2 is a Y group recited in claim 1, is a bond, or is selected from xe2x80x94C(xe2x95x90O)xe2x80x94, xe2x80x94[C(xe2x95x90O)]2xe2x80x94, xe2x80x94Oxe2x80x94, xe2x80x94NRxe2x80x94, xe2x80x94C(xe2x95x90NR)xe2x80x94, xe2x80x94S(O)1-2xe2x80x94, xe2x80x94NRC(xe2x95x90O)NRxe2x80x94, xe2x80x94NRSO2xe2x80x94, or xe2x80x94SO2NRxe2x80x94, wherein R is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, a heterocyclo or carbocyclic ring, and so forth, Rxe2x80x2 and Rxe2x80x3 may comprise substituted or unsubstituted alkylene, alkenylene, or alkynylene, and u and v may be 0-4. Additionally, the acid group xe2x80x94CO2R3 may be joined to the phenyl or pyridyl ring B with a linker such as a methylene group or replaced with other acid functional groups such as xe2x80x94SO3H, xe2x80x94P(xe2x95x90O)(OR)2, xe2x80x94SO2NHC(xe2x95x90O)R, xe2x80x94C(xe2x95x90O)NHSO2R, xe2x80x94C(xe2x95x90O)NHOH, xe2x80x94[C(xe2x95x90O)]2OR, or tetrazole, wherein R is hydrogen, alkyl, substituted alkyl, cycloalkyl, and so forth.
It should be further understood that for compounds of formula I, the linker group xe2x80x9cLxe2x80x9d is inserted into the formula I in the same direction set forth in the text. Thus, for example, if L is recited as xe2x80x94CH2xe2x80x94Yxe2x80x94, this means the xe2x80x94CH2xe2x80x94 group is attached to Z, and the Y group is attached to the C6 carbon atom i.e., to which X2 is attached, as in: 
Likewise, when Y is recited as xe2x80x94NR13C(xe2x95x90O)xe2x80x94, the carbonyl group C(xe2x95x90O) is attached to the C6 carbon atom and the nitrogen group xe2x80x94NR13xe2x80x94 is attached to Z, as in many Examples herein. Conversely, when Y is recited as xe2x80x94(CO)NR13xe2x80x94, this means the carbonyl group C(xe2x95x90O) is attached to Z and the nitrogen group xe2x80x94NR13xe2x80x94 is attached to the C6 carbon atom.
Throughout the specification, groups and substituents thereof may be chosen by one skilled in the field to provide stable moieties and compounds.
The compounds of formula I form salts which are also within the scope of this invention. Unless otherwise indicated, reference to an inventive compound is understood to include reference to salts thereof. The term xe2x80x9csalt(s)xe2x80x9d denotes acidic and/or basic salts formed with inorganic and/or organic acids and bases. In addition, the term xe2x80x9csalt(s) may include zwitterions (inner salts), e.g., when a compound of formula I contains both a basic moiety, such as an amine or a pyridine or imidazole ring, and an acidic moiety, such as a carboxylic acid. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, such as, for example, acceptable metal and amine salts in which the cation does not contribute significantly to the toxicity or biological activity of the salt. However, other salts may be useful, e.g., in isolation or purification steps which may be employed during preparation, and thus, are contemplated within the scope of the invention. Salts of the compounds of the formula I may be formed, for example, by reacting a compound of the formula I with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization.
Exemplary acid addition salts include acetates (such as those formed with acetic acid or trihaloacetic acid, for example, trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides (formed with hydrochloric acid), hydrobromides (formed with hydrogen bromide), hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates (formed with maleic acid), methanesulfonates (formed with methanesulfonic acid), 2-naphthalenesulfonates, nicotinates, nitrates, oxalates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (such as those formed with sulfuric acid), sulfonates (such as those mentioned herein), tartrates, thiocyanates, toluenesulfonates such as tosylates, undecanoates, and the like.
Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts; alkaline earth metal salts such as calcium and magnesium salts; barium, zinc, and aluminum salts; salts with organic bases (for example, organic amines) such as trialkylamines such as triethylamine, procaine, dibenzylamine, N-benzyl-xcex2-phenethylamine, 1-ephenamine, N,Nxe2x80x2-dibenzylethylene-diamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine or similar pharmaceutically acceptable amines and salts with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups may be quaternized with agents such as lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and others. Preferred salts include monohydrochloride, hydrogensulfate, methanesulfonate, phosphate or nitrate.
Prodrugs and solvates of the inventive compounds are also contemplated. The term xe2x80x9cprodrugxe2x80x9d denotes a compound which, upon administration to a subject, undergoes chemical conversion by metabolic or chemical processes to yield a compound of the formula I, and/or a salt and/or solvate thereof. Various forms of prodrugs are well known in the art. For examples of such prodrug derivatives, see:
a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, Vol.42, p. 309-396, edited by K. Widder, et al. (Acamedic Press, 1985);
b) A Textbook of Drug Design and Development, edited by Krosgaard-Larsen and H. Bundgaard, Chapter 5, xe2x80x9cDesign and Application of Prodrugs,xe2x80x9d by H. Bundgaard, p. 113-191 (1991); and
c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992), each of which is incorporated herein by reference.
Compounds containing a carboxy group can form physiologically hydrolyzable esters which serve as prodrugs by being hydrolyzed in the body to yield formula I compounds per se. For example, in compounds of formula (I), prodrugs comprise compounds wherein the upper ring substituent xe2x80x94CO2R3 is a group that will hydrolyze in the body to compounds where said substituent is xe2x80x94CO2H. Such prodrugs are preferably administered orally since hydrolysis in many instances occurs principally under the influence of the digestive enzymes. Parenteral administration may be used where the ester per se is active, or in those instances where hydrolysis occurs in the blood. Examples of physiologically hydrolyzable esters of compounds of formula I include C1-6alkylbenzyl, 4-methoxybenzyl, indanyl, phthalyl, methoxymethyl, C1-6alkanoyloxy-C1-6alkyl, e.g. acetoxymethyl, pivaloyloxymethyl or propionyloxymethyl, C1-6alkoxycarbonyloxy-C1-6alkyl, e.g. methoxycarbonyl-oxymethyl or ethoxycarbonyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl, (5-methyl-2-oxo-1,3-dioxolen-4-yl)-methyl and other well known physiologically hydrolyzable esters used, for example, in the penicillin and cephalosporin arts. Such esters may be prepared by conventional techniques known in the art.
Compounds of formula I and salts thereof may exist in their tautomeric form, in which hydrogen atoms are transposed to other parts of the molecules and the chemical bonds between the atoms of the molecules are consequently rearranged. It should be understood that the all tautomeric forms, insofar as they may exist, are included within the invention. Additionally, inventive compounds may have trans and cis isomers and may contain one or more chiral centers, therefore existing in enantiomeric and diastereomeric forms. The invention includes all such isomers, as well as mixtures of cis and trans isomers, mixtures of diastereomers and racemic mixtures of enantiomers (optical isomers). When no specific mention is made of the configuration (cis, trans or R or S) of a compound (or of an asymmetric carbon), then any one of the isomers or a mixture of more than one isomer is intended. The processes for preparation can use racemates, enantiomers or diastereomers as starting materials. When enantiomeric or diastereomeric products are prepared, they can be separated by conventional methods for example, chromatographic or fractional crystallization.
The compounds of the instant invention may, for example, be in the free or hydrate form, and may be obtained by methods exemplified by the following descriptions.
Preferred compounds are those having the formula (I), 
and pharmaceutically-acceptable salts, prodrugs, or solvates thereof, in which:
W is selected from xe2x80x94C(xe2x95x90O)NR4R5, xe2x80x94OR6, optionally-substituted heterocycle, substituted alkyl, alkenyl, and substituted alkenyl;
ring B is phenyl;
X2 is N, CH, or C, provided that X2 is C when R1 and R2 join to form a fully unsaturated ring;
L is xe2x80x94(CH2)sxe2x80x94Yxe2x80x94;
Y is selected from xe2x80x94C(xe2x95x90O), xe2x80x94NHxe2x80x94C(xe2x95x90O)xe2x80x94, xe2x80x94NHxe2x80x94CH2xe2x80x94, and xe2x80x94CH2xe2x80x94CH2xe2x80x94;
Z is selected from 
R1 and R2 (i) are independently selected from hydrogen, lower alkyl, aryl and arylalkyl; or (ii) are taken together to form an aryl, heteroaryl, cycloalkyl, or heterocyclo; wherein when R1 and R2 individually or together form a heteroaryl, aryl, heterocyclo or cycloalkyl, said cyclic group is optionally substituted with up to two R26;
R3 is hydrogen, alkyl, substituted alkyl, or alkyl substituted with xe2x80x94OC(xe2x95x90O)R24 or xe2x80x94OC(xe2x95x90O)OR24, wherein R24 is alkyl, substituted alkyl, or cycloalkyl;
R4 is hydrogen or lower alkyl;
R5 is hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, heterocyclo or heteroaryl;
R6 is selected from C1-6alkyl, more preferably C2-6alkyl, phenyl, and benzyl;
R8 and R26 (i) are at each occurrence independently selected from hydrogen, OR30, NR31R32, alkyl, alkenyl, substituted alkyl, substituted alkenyl, halogen, haloalkyl, haloalkoxy, cyano, nitro, alkylthio, xe2x80x94C(xe2x95x90O)H, acyl, xe2x80x94CO2H, alkoxycarbonyl, sulfonamido, sulfonyl, and phenyl, or (ii) two of R8 and/or two of R26 may be taken together to form a fused benzo ring, a fused heteroaryl, or a fused heterocyclo other than a five or six membered heterocyclo having as its heteroatoms two oxygen atoms, provided further that when two R26 form a fused benzo ring, then Z is not phenyl substituted in the para position with cyano or a five-membered heterocycle or heteroaryl;
R20 and R21 are independently selected from hydrogen, halogen, xe2x80x94C(xe2x95x90O)NH2, xe2x80x94C(xe2x95x90O)C1-4alkyl, xe2x80x94NH2, xe2x80x94NHC1-4alkyl, xe2x80x94Sxe2x80x94C1-4alkyl, xe2x80x94Oxe2x80x94C1-4alkyl, C1-4alkyl, C1-4alkyl substituted with NH2, and five or six membered heterocyclo or heteroaryl;
R30 at each occurrence is selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, and phenyl;
R31 and R32 at each occurrence are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, and cycloalkyl;
m and n are independently 0, 1 or 2; and
s is 0, 1 or 2.
In compounds of formula I, the group W is preferably xe2x80x94C(xe2x95x90O)NR4R5 and the groups Zxe2x80x94Lxe2x80x94 taken together are preferably selected from: 
More preferred compounds are those compounds having the formulae: 
in which:
X3 is CH or N;
R3 is hydrogen, lower alkyl, or lower alkyl substituted with one of OC(xe2x95x90O)R24 and OC(xe2x95x90O)Oxe2x80x94R24, wherein R24 is alkyl or cycloalkyl;
R4 is hydrogen or lower alkyl;
R5 is C1-6alkyl, xe2x80x94CH(CH2OH)C(CH3)3, or C1-2alkyl substituted with C5-6cycloalkylene;
either (a) s is 0 and Z is selected from 
(b) s is 1 and Z is selected from 
R26 is C2-6straight or branched alkenyl, xe2x80x94OR30 or xe2x80x94NR31R32, and R27 is hydrogen, or R26 and R27 together form a fused benzo ring;
R30 is C1-5 straight or branched chain alkyl, C2-6straight or branched alkenyl, C3-5cycloalkyl, or C1-4 straight or branched chain alkyl substituted with one to two of halogen, lower alkoxy, and C3-5cycloalkyl;
R31 and R32 are selected from hydrogen and lower alkyl.
Most preferred are compounds having the formula: 
in which
R3 is hydrogen, lower alkyl, or lower alkyl substituted with xe2x80x94OC(xe2x95x90O)R24 or xe2x80x94OC(xe2x95x90O)OR24, wherein R24 is alkyl or cycloalkyl;
Z is selected from: 
R25 is hydrogen or hydroxymethyl; and
R26 is C1-3alkoxy or NH(C1-4alkyl).
The compounds of the invention may be prepared by the exemplary processes described in the following Schemes A through D. Methods for making intermediates including appropriately-protected amine-coupling components are shown in Schemes E through G and I through X, and Scheme H shows a method for making an unprotected amine-coupling component. These amines may be coupled to substrates to make compounds of formula I and deprotected, when necessary or desired, as shown in Schemes A-D and the Examples. Exemplary reagents and procedures for these reactions appear hereinafter and in the working Examples. Starting materials are commercially available or can be readily prepared by one of ordinary skill in the art as shown herein or as described in the literature. For all of the schemes, the groups R1-R27, W, X, Z, r, s etc., are as described herein for a compound of formula I, unless otherwise indicated. 
Compounds of formula Ia can be made by reacting acid 1 with an amine having the desired group Z, i.e., Zxe2x80x94NHR13. The 2-position acid group is suitably protected (Pxe2x80x2), and the reaction is carried out in the presence of coupling reagent(s) such as DCC/HOBT/DMAP, EDC/DMAP, or DIC/HOAT to afford the corresponding amide compound. The group Pxe2x80x2 optionally may be deprotected to afford the compound of formula Ia wherein R3 is hydrogen, or the group Pxe2x80x2 may be retained wherein Pxe2x80x2 comprises the desired group R3. Alternatively, the group Pxe2x80x2 may be deprotected to afford the group CO2H, with the group CO2H then converted to another desired R3 group. To illustrate, the compound having the acid group CO2H may be reacted with a halide having the desired R3 group, i.e., Xxe2x80x94R3 where X is Cl, Br, or I, in the presence of base, or the acid compound may be coupled with an alcohol such as R3OH in a coupling reagent. It may be necessary or desired to protect additional functional groups besides the 2-position acid before performing the coupling reaction, as one skilled in the field will appreciate. Those additional protecting groups can be removed after the coupling using appropriate deprotecting conditions. Preparation of acids 1, wherein R1 and R2 together form an unsaturated carbocyclic or heterocyclic ring, is described in WO 99/041231, incorporated herein by reference, and described in the Examples that appear hereinafter. 
Similar to Scheme A, the aldehyde 2, wherein the 2-position acid group is suitably protected (Pxe2x80x2), can be coupled with an amine Zxe2x80x94NHR13 in the presence of a reducing reagent such as sodium triacetoxyborohydride, to afford the corresponding amine compound having the group CO2Pxe2x80x2. Upon optional deprotection of the group Pxe2x80x2, and optionally further reaction with, for example, a halide Xxe2x80x94R3 or alcohol R3OH as described in Scheme A, the compound of formula Ib is provided, having the desired group R3. Also as in Scheme A, it may be necessary or desired to protect additional functional groups besides the 2-position acid before performing the coupling reaction. Those additional protecting groups can be removed using appropriate deprotecting conditions. Preparation of aldehydes 2, wherein R1 and R2 together form an unsaturated carbocyclic or heterocyclic ring, is described in WO 99/041231, incorporated herein, and further shown in the Examples hereinafter. 
Aryl fluoride 3a is reacted with amine 4 in DMSO in the presence of a base such as DIEA to afford intermediate 5. Alternatively, triflate 3b is reacted with amine 4 in the presence of a suitable palladium reagent to afford intermediate 5. Selective deprotection of the Pxe2x80x3 group of compound 5 affords acid 6. Acid 6 is reacted with an appropriate amine in the presence of suitable coupling reagents. Compounds having the desired group R3 are obtained as described in Scheme A, i.e., by optional deprotection, further reaction or coupling, to afford compounds of formula IIa, above. 
Aryl fluoride 3a is reacted with amine 7 in DMSO in the presence of a base such as DIEA to afford compound 8, where R1 is defined as above except where R1 and R2 form a ring, the ring is a heterocyclo. Selective deprotection of the Pxe2x80x3 group affords acid 9. Reaction of acid 9 with an amine Zxe2x80x94NHR13 in the presence of coupling reagent(s) such as DCC/HOBT/DMAP, EDC/DMAP, or DIC/HOAT affords the corresponding amide compound. Compounds having the desired group R3 are obtained as described in Scheme A, i.e., by optional deprotection, further reaction or coupling, to afford compounds of formula IIb, above. 
Compound 10 was prepared according to J. Med. Chem., Vol. 42 (1999), at pp. 3510-3519, from 2-methyl-4-nitroaniline. A mixture of compound 10 and 1-(1,1-dimethylethoxy)-N,N,Nxe2x80x2,Nxe2x80x2-tetramethyl-methanediamine in dry DMF (10 mL) was stirred at 70xc2x0 C. for 2 h under N2. After cooling to rt, the reaction mixture was treated with hexane, and the solid was collected by filtration and washed with hexane to give compound 11 as black crystals. Compound 11 was converted to compound 13 in two alternate ways.
In one approach, compound 11 was converted to 13 by adding 1N LiHMDS to a solution of 11 in dry THF under N2. The reaction mixture was stirred at 65xc2x0 C. for 2 h. After cooling to rt, 12 N HCl was added and the reaction mixture stirred at 50xc2x0 C. for 1 h. After cooling to rt, the mixture was neutralized with sat""d NaHCO3, the product extracted with EtOAc, and the organic layer washed with water and sat""d NaCl. The product was concentrated and purified to give compound 13 as a yellow solid.
Alternatively, compound 11 was converted to 13 by first mixing compound 11 and 2,4-dimethoxylbenzylamine in DMF and stirring the mixture at 140xc2x0 C. for 3 h. The solvent was removed by vacuum distillation and residue treated with EtOAc. The orange solid was collected by filtration and washed with hexane to give compound 12. To a solution of compound 12 in anisole was added TFA. The reaction mixture was stirred at 90xc2x0 C. for 1 h and the solvent removed under reduced pressure. The residue was treated with sat""d NaHCO3 (30 mL) and the product collected by filtration and washed with water to afford compound 13.
Compound 13 (366 mg, 1.93 mmol) and 2,4-dimethoxybenzaldehyde were heated for 16 h at 125-130xc2x0 C. with a stream of nitrogen passing in and out of the reaction flask, and sampling of the reaction mixture at 80xc2x0 C. indicated conversion to compound 14.
To a solution of 14 and 2,4-dimethoxybenzaldehyde above in THF was added sodium triacetoxyborohydride. The reaction was stirred for 22 h and additional sodium triacetoxyborohydride (1.23 g, 5.8 mmol) was added. After 40 h, the reaction was concentrated to an oil which was taken up in EtOAc, water, and dilute sodium bicarbonate. The EtOAc was washed with water (3xc3x97), dried (sodium sulfate), and concentrated to an oily residue, which was chromatographed to give 140 mg of compound 15a as a glassy residue and 228 mg of compound 15b as an amorphous solid.
Hydrogenation of compound 15b in EtOAc and MeOH in the presence 10% Pd/C for 1 h at one atmosphere afforded compound 16 as an amorphous solid. Compound 16 was coupled to a substrate and deprotected to produce compounds of formula I. 
A mixture of compound 13 and di-t-butyl dicarbonate in dry THF was refluxed under N2 for 3 h. The mixture was concentrated and the residue purified by flash chromatography eluting with EtOAc/Hexane (1:3) to give compound 17 as a white solid. Compound 17 and Pd/C (10%) in MeOH/dioxane was hydrogenated (balloon with H2) for 3.5 h. Filtration and concentration yielded 18 as a brown foam (59 mg, 83%), which was used in Examples hereinafter as a protected amine-coupling component to make compounds of formula I. 
Compound 20 was synthesized from compound 19 following the procedure described in Osborn, et al., J.Chem. Soc. (1956), at 4191, and compounds 21a and 21b were prepared according to Poradowska et al., Synthesis, (1975), at p. 732. Compound 21a and phthalic acid anhydride 22 were powdered and mixed well. Heating the mixture for 2 h at 130 to 150xc2x0 C. and finally 2 min to 220xc2x0 C. finished the reaction. The cooled solid material of crude compound 23 was powdered and washed with ether/DCM (10:1) and dried yielding compound 23 as a beige powder.
Compound 23 and MCPA (Aldrich,xcx9c77%) were dissolved in DCE and stirred for 24 h. The resulting suspension was diluted with 50 ml ether and the crude product filtered, washed with ether, dried and purified to yield compound 24 as a light yellow powder. Compound 24 and POCl3 were heated for 12 h to 90xc2x0 C. Excess POCl3 was removed in vacuo and the residue stirred with ice water/DCM for 10 min. The organic layer was dried over Na2SO4 and concentrated. The oily residue was purified to yield compound 25. POCl3 was removed by dissolving the material in DCM and stirring with N -diisopropylaminomethyl polystyrol. Filtration, concentration, and purification gave compound 25 as off-white needles.
Compound 25 and N-methylhydrazine were stirred in DCM to produce compound 26. Compound 26 and 2,4-dimethoxybenzylamine were heated to 110 to 120xc2x0 C. and stirred to produce an oily crude material which was purified to give protected amine-coupling component 27 as a beige foam. 
Compound 26 from Scheme G and condensed N-methylamine were heated to 100xc2x0 C. for 24 h. Cooling, removal of the excess N-methylamine, and purification gave unprotected amine-coupling component 28 as an off-white solid. 
Following the procedure described in Scheme G, compound 29 was prepared from compound 19 and phthalic acid anhydride 22; compound 30 was prepared from compound 29 and MCPA; compound 31 was prepared from 30 and POCl3; compound 32 was prepared from compound 31 and methylhydrazine; and protected amine-coupling component 33 was prepared from 32 and dimethoxybenzylamine. 
Compound 34 and bis-protected isothiourea 35 were suspended in MeOH and refluxed for 5 days. After the second day, n-BuOH was added and CH3SH blown out with N2. The reflux temperature was set to 100xc2x0 C. After 3 more days refluxing, compound 36 crystallized and the reaction was completed. The mixture was cooled to 50xc2x0 C., filtered, and the gray filter cake was washed with MeOH and recrystallized from DMF/MeOH to give compound 36 in the form of grey fine crystals.
Compound 36 was suspended in AcOH with stirring. Zn powder was added. After 1 hr, the reaction mixture was filtered, the filtrate concentrated, water was added, and then filtration, washing of the filter residue, and drying gave Cbz-protected amine coupling component 37 as a purple powder. 
Cyanogen bromide was added to a flask charged with compound 34, water and EtOH. After 12 h, the reaction mixture was filtered, the filtrate was basified to pH=9 using conc. NH4OH, the solution was conc. to one third volume, and H2O was added. After 1 h at 4xc2x0 C., the solid was filtered and dried under vacuum to give compound 38.
A solution of Boc anhydride in THF was added to a cold (0xc2x0 C.) solution of compound 38 in THF (90 mL). DMAP was added, and the reaction mixture was stirred at rt. After 30 min, the solution was concentrated, the residue was dissolved in DCM, and then the solution was washed with 2% aq. NH4Cl and sat. NaCl, dried (MgSO4), and conc. to give compound 39.
MeOH and EtOAc (3:1) was added to compound 39. 10% Pd/C was added and a H2 atmosphere introduced via balloon. After 12 h, the reaction mixture was filtered, the filtrate was conc., and the residue was placed under vacuum to give Boc-protected amine coupling component 40. 
Compound 41 was dissolved in pyridine and while stirring, 2.55 g (25 mM) acetic acid anhydride was added over 15 min. Stirring continued for 24 h. The product was concentrated in vacuo and the resulting oil taken up with DCM/water. The pH was adjusted to 3.0 with citric acid. The phases were separated and the aqueous layer washed two more times with 50 ml DCM each time. The combined organic layer was washed with brine, dried (MgSO4), and concentrated to give compound 42.
Compound 42 and CDI were dissolved in 40 ml THF. After 30 min stirring, the solution was slowly added at 0xc2x0 C. to 150 ml sat""d solution of NH3 gas in THF. After stirring for 24 h at rt, the reaction mixture was filtered and the filtrate concentrated. The oily residue was dissolved in 30 ml MeOH from which after several minutes 43a crystallized. The mother liquor contained a mixture of 43a and 43b. Refluxing for 4 h in the presense of TosOH led to a complete cyclizsation of 43b to 43a.
Compound 43a and 760 mg (1.90 mM) Laweson reagent were suspended in 70 ml xylene and refluxed at 140xc2x0 C. for 3 h. After cooling to rt, compound 44 crystallized out of the solution.
Compound 44 and 3 ml 1.0 N NaOH were dissolved in 15 ml DMF. To the stirred solution was added 166 ul (3.10 mM) CH3. After 5 min, compound 45 crystallized, 50 ml water/EtOAc was added, and the aq. layer was extracted two times with 20 ml EtOAc. The combined organic phase was washed with brine, dried (Na2SO4), and the product 45 concentrated and purified.
Compound 45 and 2,4-dimethoxybenzylamine were dissolved in 10 ml toluene and refluxed for 2.5 h. After adding 30 ml ether to the cooled solution, filtration and drying, compound 46 was obtained as yellow crystals.
Compound 46 was dissolved in 10 ml AcOH and while stirring, 200 mg (3.06 mM) Zn powder was added. After 30 min, filtered from excess Zn, washed with 5 ml AcOH, and concentrated in vacuo yield an oily residue of crude 27 AcOH salt. This material was taken up with 20 ml water and the pH adjusted to 10 with Na2CO3 solution, followed by extraction 3 times with 15 ml EtOAc. The combined organic layer was washed with brine, dried (Na2SO4), and concentrated yielding protected amine-coupling component 47 in the form of a white foamy material. 
Nitro and carboxylic acid starting materials (e.g. 48) were dissolved in DCM and N,N-DMF (10:1). 1,1xe2x80x2-carbonyldiimidazole (1.2 equiv) was added, and the reaction stirred at rt for 5 h. Ammonium hydroxide (2 equiv) was then added. After stirring overnight at rt, the reaction was concentrated, washed with base and extracted with EtOAc to yield compound 49. Compound 49 was then hydrogenated at 40 psi on the PARR shaker in the presence of Pd/C catalyst. Filtration and concentration yielded the appropriate Z-amine coupling component 50. This same or similar method was used to make 
To a solution of compound 51 in 20 mL of pyridine was added toluenesulfonyl chloride. The solution was stirred for 18 h at 80xc2x0 C. and cooled to rt and concentrated. The precipitate was taken up with water, filtered, and washed with water. The solid was then crystallized from EtOAc to give compound 52 as white needle crystals.
To a solution of compound 52 in 20 mL of N,N-DMF was added 0.79 g (6.1 mmol) of DIPEA and 1.13 g (6.1 mmol) of iodoacetamide at rt. The solution was stirred for 24 h and then poured into 100 mL of water and stirred for 1 h. The solid was collected and dried under vacuum to yield compound 53. Compound 53 was taken up with 20 mL DCM and 1.2 g (6.1 mmol) of trifluoroacetic anhydride was added at rt. The resulting solution was stirred for 5 h at rt and concentrated. The residue was taken up with EtOAc and washed with saturated sodium bicarbonate. The organic layer was dried over MgSO4 and concentrated to give compound 54 as a white solid. MS, m/z (M+1)+=289.
The resulting compound 54 was dissolved in EtOAc and Pd/C catalyst was added. The mixture was placed on the PARR shaker at 40 psi for 2 h. Filtering off the catalyst yielded the desired compound 55 in 80% yield. 
To a solution of 4-bromo-3-fluorotoluene (2.0 g, 10.58 mmol) in CCl4 (40 ml) at RT was added NBS (2.0 g). The reaction was heated to reflux and benzoylperoxide (128 mg, 0.53 mmol) was added three times (total 384 mg) in 30 minute intervals. The reaction was cooled to RT, diluted with DCM (40 ml) and washed with sat. NaHCO3 (2xc3x97). The organics were dried over MgSO4, filtered and concentrated to isolated 56 (2.8 g crude).
To a solution of compound 56 (2.8 g, xcx9c10.4 mmol) in DMF (45 ml) under nitrogen at RT was added BOC2NH (3.4 g, 15.7 mmol) followed by KOtBu (1.76 g, 15.7 mmol). After a mild exotherm, the reaction was stirred at rt for 72 hr. The reaction was diluted with EtOAc (200 ml) and washed with 1N HCl, water, sat. NaHCO3 and brine. Organics were dried over MgSO4, filtered and concentrated. Purification by flash chromatography (silica gel, 2%-15% EtOAc in hexane) provided compound 57 (1.56 g). MS (M+Na)+=426, 428 (Br isotopic pattern).
To a solution of 57 (1.56 g, 3.84 mmol) in nitrogen degassed DMF (1% water, 20 ml) was added Pd(dba)3 (70.3 mg, 0.077 mmol), DPPF (95.9 mg, 0.173 mmol) and Zn(CN)2 (315 mg, 2.69 mmol). The reaction mixture was degassed with nitrogen for 30 minutes, sealed and heated at 110xc2x0 C. for 20 hours. The reaction was diluted with EtOAc (100 ml) and filtered through a plug of celite which was then washed with EtOAc (2xc3x9750 ml). The eluent was then placed in a separatory funnel and washed with water (3xc3x97150 ml). The water layers were back extracted in the order generated. The combined EtOAc extracts were dried over MgSO4, filtered and concentrated. Purification by flash chromatography (silica gel, 0 to 15% EtOAc in hexane) provided 58a (0.71 g) and 58b (0.45 g).
To a solution of acetohydroxamic acid (135 mg, 1.8 mmol) in DMF (5 ml) at RT was added KOtBu (1.0M THF, 1.8 ml, 1.8 mmol). A gelatinous suspension formed which was aggitated until well mixed. The suspension was allowed to set at RT for 5 minutes and then a solution of 58b (450 mg, 1.8 mmol) in DMF (15 ml) was added. The reaction was aggitated at RT for 20 hours. The reaction mixture was then diluted with EtOAc (150 ml) and washed with water (2xc3x97) and brine (1xc3x97). The organics were dried over MgSO4, filtered, and concentrated. Crystallization from CH2Cl2/hexane gave compound 59 (240 mg). MS (M+H)+=264
Compound 59 (240 mg) was treated with 10%TFA/CH2Cl2 (5 ml) at RT for 3 hours. Solvents were removed and the residue was taken up with MeOH and added to a plug of Dowex 50W-X2 (H form, 10 g) resin. The resin captured amine was washed with MeOH, DCM and CH3CN. Elution with 2M NH3 in MeOH (60 ml) followed by concentration gave Z-amine coupling component 60. (170 mg). 
To a solution of 3-amino-5-nitrobenzisoxazole (200 mg, 1.12 mmol, lit. WO/0027627) in DCM (5 ml) was added BOC2O (536 mg, 2.46 mmol) followed by DMAP (20 mg). The reaction mixture was stirred overnight. Solvent was removed and purification by flash chromatography (silica gel, CH2Cl2) gave a mixture of compounds 61a and 61b (350 mg combined).
The mixture of 61a and 61b (307 mg combined) was taken up in EtOH (10 ml) and treated with SnCl2.2H2O (751 mg). The reaction was heated to 70xc2x0 C. for 1.5 hours. The reaction was diluted with EtOAc (75 ml), water (50 ml) and sat. NaHCO3 (25 ml). The layers were mixed and allowed to separate. The organic layer was dried over MgSO4, filtered, and concentrated. Purification by RP Prep HPLC provided BOC-protected amine-coupling component 62. (101 mg). MS (M+H)+=250.
The inventive compounds are inhibitors of the activated coagulation serine proteases known as Factor VIIa, Factor IXa, Factor Xa, Factor XIa, and thrombin and also inhibit other serine proteases, such as trypsin, tryptase, and urokinase. Thus, the compounds are useful for treating or preventing those processes, which involve the production or action of Factor VIIa, Factor IXa, Factor Xa, Factor XIa, thrombin, trypsin, and/or tryptase. In view of their urokinase inhibitory activity, they are useful as metastasis inhibitors in treating cancer. As used herein with reference to the utilities described below, the term xe2x80x9ctreatingxe2x80x9d or xe2x80x9ctreatmentxe2x80x9d encompasses prevention, partial alleviation, or cure of the disease or disorder.
In view of their above-referenced serine protease inhibitory activity, the inventive compounds are useful in treating consequences of atherosclerotic plaque rupture including cardiovascular diseases associated with the activation of the coagulation cascade in thrombotic or thrombophilic states. Such diseases include arterial thrombosis, coronary artery disease, acute coronary syndromes, myocardial infarction, unstable angina, ischemia resulting from vascular occlusion cerebral infarction, stroke and related cerebral vascular diseases (including cerebrovascular accident and transient ischemic attack). Additionally, the compounds are useful in treating or preventing formation of atherosclerotic plaques, transplant atherosclerosis, peripheral arterial disease and intermittent claudication. In addition, the compounds can be used to prevent restenosis following arterial injury induced endogenously (by rupture of an atherosclerotic plaque), or exogenously (by invasive cardiological procedures such as vessel wall injury resulting from angioplasty).
In addition, the inventive compounds are useful in preventing venous thrombosis, coagulation syndromes, deep vein thrombosis (DVT), disseminated intravascular coagulopathy, Kasabach-Merritt syndrome, pulmonary embolism, cerebral thrombosis, atrial fibrillation, and cerebral embolism. The compounds are useful in treating peripheral arterial occlusion, thromboembolic complications of surgery (such as hip replacement, endarterectomy, introduction of artificial heart valves, vascular grafts, and mechanical organs), implantation or transplantation of organ, tissue or cells, and thromboembolic complications of medications (such as oral contraceptives, hormone replacement, and heparin, e.g., for treating heparin-induced thrombocytopenia). The inventive compounds are useful in preventing thrombosis associated with artificial heart valves, stents, and ventricular enlargement including dilated cardiac myopathy and heart failure. The compounds are also useful in treating thrombosis due to confinement (i.e. immobilization, hospitalization, bed rest etc.).
These compounds are also useful in preventing thrombosis and complications in patients genetically predisposed to arterial thrombosis or venous thrombosis (including activated protein C resistance, FVleiden, Prothrombin 20210, elevated coagulation factors FVII, FVIII, FIX, FX, FXI, prothrombin, TAFI and fibrinogen), elevated levels of homocystine, and deficient levels of antithrombin, protein C, and protein S. The inventive compounds may be used for treating heparin-intolerant patients, including those with congenital and acquired antithrombin III deficiencies, heparin-induced thrombocytopenia, and those with high levels of polymorphonuclear granulocyte elastase.
The present compounds may also be used to inhibit blood coagulation in connection with the preparation, storage, fractionation, or use of whole blood. For example, the compounds may be used to maintain whole and fractionated blood in the fluid phase such as required for analytical and biological testing, e.g., for ex vivo platelet and other cell function studies, bioanalytical procedures, and quantitation of blood-containing components. The compounds may be used as anticoagulants in extracorpeal blood circuits, such as those necessary in dialysis and surgery (such as coronary artery bypass surgery); for maintaining blood vessel patency in patients undergoing transluminal coronary angioplasty, vascular surgery including bypass grafting, arterial reconstruction, atherectomy, vascular graft and stent patency, tumor cell metastasis, and organ, tissue, or cell implantation and transplantation.
In view of their tryptase inhibitory activity, the inventive compounds are useful as anti-inflammatory agents, in treating chronic asthma, allergic rhinitis, inflammatory bowel disease, psoriasis, conjunctivitis, atopic dermatitis, pancreatis, rheumatoid arthritis, osteoarthritis, septic shock, and chronic inflammatory joint diseases, diseases of joint cartilage destruction, and/or vascular damage due to bacterial and/or viral infections. Additionally, the inventive compounds may be useful for treating diabetic retinopathy or motor neuron diseases such as amyotrophic lateral sclerosis, progressive muscular atrophy, and primary lateral sclerosis. Additionally, the inventive compounds may be useful for tissue remodeling diseases and for treating plaque instability and sequelli. In addition, these compounds may be useful for treating fibrotic diseases and conditions, for example, fibrosis, scleroderma, pulmonary fibrosis, liver cirrhosis, myocardial fibrosis, neurofibromas, and hypertrophic scars.
In addition, the compounds of the present invention are useful in treating cancer and preventing the prothrombotic complications of cancer. In view of their metastasis inhibition activity, the compounds are useful in treating tumor growth, as an adjunct to chemotherapy, and for treating diseases involving metastases including, but not limited to cancer, more particularly, cancer of the lung, prostate, colon, breast, ovaries, and bone. These compounds may also be useful in preventing angiogenesis.
The inventive compounds may also be used in combination with other antithrombotic or anticoagulant drugs such as thrombin inhibitors, platelet aggregation inhibitors such as aspirin, clopidogrel, ticlopidine or CS-747, warfarin, low molecular weight heparins (such as LOVENOX), GPIIb/GPIIIa blockers, PAI-1 inhibitors such as XR-330 and T-686, inhibitors of xcex1-2-antiplasmin such as anti-xcex1-2-antiplasmin antibody and thromboxane receptor antagonists (such as ifetroban), prostacyclin mimetics, phosphodiesterase (PDE) inhibitors, such as dipyridamole or cilostazol, PDE inhibitors in combination with thromboxane receptor antagonists/thromboxane A synthetase inhibitors (such as picotamide), serotonin-2-receptor antagonists (such as ketanserin), fibrinogen receptor antagonists, hypolipidemic agents, such as HMG-CoA reductase inhibitors, e.g., pravastatin, simvastatin, atorvastatin, fluvastatin, cerivastatin, AZ4522, itavastatin (Nissan/Kowa), and compounds disclosed in U.S. provisional applications No. 60/211,594 filed Jun. 15, 2000, and No. 60/211,595 filed Jun. 15, 2000; microsomal triglyceride transport protein inhibitors (such as disclosed in U.S. Pat. Nos. 5,739,135, 5,712,279 and 5,760,246), antihypertensive agents such as angiotensin-converting enzyme inhibitors (e.g., captopril, lisinopril or fosinopril); angiotensin-II receptor antagonists (e.g., irbesartan, losartan or valsartan); and/or ACE/NEP inhibitors (e.g., omapatrilat and gemopatrilat); xcex2-blockers (such as propranolol, nadolol and carvedilol), PDE inhibitors in combination with aspirin, ifetroban, picotamide, ketanserin, or clopidogrel and the like. The inventive compounds are also useful in combination with anti-arrhythmic agents such as for atrial fibrillation, for example, amiodarone or dofetilide.
The inventive compounds may be used in combination with prothrombolytic agents, such as tissue plasminogen activator (natural or recombinant), streptokinase, reteplase, activase, lanoteplase, urokinase, prourokinase, anisolated streptokinase plasminogen activator complex (ASPAC), animal salivary gland plasminogen activators, and the like.
The inventive compounds may also be used in combination with xcex2-adrenergic agonists such as albuterol, terbutaline, formoterol, salmeterol, bitolterol, pilbuterol, or fenoterol; anticholinergics such as ipratropium bromide; anti-inflammatory cortiocosteroids such as beclomethasone, triamcinolone, budesonide, fluticasone, flunisolide or dexamethasone; and anti-inflammatory agents such as cromolyn, nedocromil, theophylline, zileuton, zafirlukast, monteleukast and pranleukast.
The inventive compounds may also be useful in combination with other anticancer strategies and chemotherapies such as taxol and/or cisplatin.
The compounds may act synergistically with one or more of the above agents. For example, the inventive compounds may act synergistically with the above agents to prevent reocclusion following a successful thrombolytic therapy and/or reduce the time to reperfusion. Thus, reduced doses of thrombolytic agent(s) may be used, therefore minimizing potential hemorrhagic side effects.
The compounds of formula I may be administered by any means suitable for the condition to be treated, which may depend on the need for site-specific treatment or quantity of drug to be delivered. Systematic treatment is typically preferred for cancerous conditions, although other modes of delivery are contemplated. The compounds may be delivered orally, such as in the form of tablets, capsules, granules, powders, or liquid formulations including syrups; sublingually; bucally; transdermally; parenterally, such as by subcutaneous, intravenous, intramuscular or intrasternal injection or infusion (e.g., as sterile injectable aqueous or non-aqueous solutions or suspensions); nasally such as by inhalation spray; rectally such as in the form of suppositories, or in the form of liposome particles. Dosage unit formulations containing non-toxic, pharmaceutically acceptable vehicles or diluents may be administered. The compounds may be administered in a form suitable for immediate release or extended release. Immediate release or extended release may be achieved with suitable pharmaceutical compositions or, particularly in the case of extended release, with devices such as subcutaneous implants or osmotic pumps.
Exemplary compositions for oral administration include suspensions which may contain, for example, microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners or flavoring agents such as those known in the art; and immediate release tablets which may contain, for example, microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and/or lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants such as those known in the art. The inventive compounds may be orally delivered by sublingual and/or buccal administration, e.g., with molded, compressed, or freeze-dried tablets. Exemplary compositions may include fast-dissolving diluents such as mannitol, lactose, sucrose, and/or cyclodextrins. Also included in such formulations may be high molecular weight excipients such as celluloses (AVICEL(copyright)) or polyethylene glycols (PEG); an excipient to aid mucosal adhesion such as hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC), sodium carboxymethyl cellulose (SCMC), and/or maleic anhydride copolymer (e.g., GANTREZ(copyright)); and agents to control release such as polyacrylic copolymer (e.g., CARBOPOL 934(copyright)). Lubricants, glidants, flavors, coloring agents and stabilizers may also be added for ease of fabrication and use.
Exemplary compositions for nasal aerosol or inhalation administration include solutions which may contain, for example, benzyl alcohol or other suitable preservatives, absorption promoters to enhance absorption and/or bioavailability, and/or other solubilizing or dispersing agents such as those known in the art.
Exemplary compositions for parenteral administration include injectable solutions or suspensions which may contain, for example, suitable non-toxic, parenterally acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer""s solution, an isotonic sodium chloride solution, or other suitable dispersing or wetting and suspending agents, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.
Exemplary compositions for rectal administration include suppositories which may contain, for example, suitable non-irritating excipients, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures but liquefy and/or dissolve in the rectal cavity to release the drug.
The effective amount of a compound of the present invention may be determined by one of ordinary skill in the art. The specific dose level and frequency of dosage for any particular subject may vary and will depend upon a variety of factors, including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the species, age, body weight, general health, sex and diet of the subject, the mode and time of administration, rate of excretion, drug combination, and severity of the particular condition. An exemplary effective amount of compounds of formula I may be within the dosage range of about 0.1 to about 100 mg/kg, preferably about 0.2 to about 50 mg/kg and more preferably about 0.5 to about 25 mg/kg (or from about 1 to about 2500 mg, preferably from about 5 to about 2000 mg) on a regimen in single or 2 to 4 divided daily doses.
Enzyme Assays
Compound was prepared as a 5 mM stock in DMSO, diluted further in DMSO and added directly to the assays. The DMSO concentration for all these studies was less than 1% and compared to DMSO vehicle controls.
Human Factor VIIa was obtained from Enzyme Research Labs (Cat.# HFVIIA 1640). Human recombinant tissue factor (INNOVIN from Dade Behring Cat.# B4212-100; xe2x80x9c20 ml vialxe2x80x9d) was diluted with 8 ml of H2O per vial and diluted further 1:30 into the 302 xcexcl final assay volume. Tissue factor activated FVIIa enzymatic activity was measured in a buffer containing 150 mM NaCl, 5mM CaCl2, 1 mM CHAPS and 1 mg/ml PEG 6000 (pH 7.4) with 1 nM FVIIa and 100 xcexcM D-Ile-Pro-Arg-AFC (Enzyme Systems Products, Km greater than 200 xcexcM) 0.66% DMSO. The assay (302 xcexcl total volume) was incubated at RT for 2 hr prior to reading fluorometric signal (Ex 405/Em 535) using a Victor 2 (Wallac) fluorescent plate reader.
Human Factor IXa (American Diagnostica #449b) enzymatic activity was measured in a buffer containing 50 mM Tris, 100 mM CaCl2, 5 mM CaCl2, 33% ethylene glycol at pH 7.5 using 96-well microtiter plates (Nunc #439454). The enzyme was incubated with the inhibitor at RT for three minutes prior to starting the reaction with 500 uM Spectrozyme FIXa (American Diagnostica #299). The Km for this substrate is estimated by American Diagnostica to be 1.3 mM. Time dependent optical density change was followed at 405 nm using a kinetic microplate read (Molecular Devices Spectramax Plus) at RT. Enzyme activity in the presence of inhibitor was expressed as fraction of a DMSO-containing control and curve fit to the equation: activity=control activity/(1+[I]/IC50) using Excel Fit.
Human FXa (Calbiochem #233526) enzymatic activity was measured in a buffer containing 0.145 M NaCl, 0.005 M KCl, 1 mg/ml Polyethylene Glycol (PEG-8000), 0.030 M HEPES (pH 7.4) using 96-well microtiter plates (Nunc Immuno #439454). The enzyme was incubated with the inhibitor at RT for three minutes prior to starting the reaction with 100 xcexcM S-2222 (phenyl-Ile-Glu-Gly-Arg-pNA, Km=137 xcexcM). The Km for this and other substrates was determined experimentally by measuring the enzyme activity at different substrate concentrations and curve fitting the data using Kaleidagraph V. Time-dependent optical density change was followed at 405 nm using a kinetic microplate reader (Molecular Devices UVmax) at RT. Enzyme activity in the presence of inhibitor was expressed as fraction of a DMSO-containing control and curve fit to the equation: activity=control activity/(1+[I]/IC50) using Excel Fit.
Recombinant urokinase (Abbott Labs, Abbokinase) was assayed in the same buffer as FXa, but the reactions were started with 100 xcexcM S-2444 (L-pyroGlu-Gly-Arg-pNA, Km=31 xcexcM). Human xcex1-thrombin (Sigma) was measured as for FXa except that the reaction was started with 10 xcexcM S-2238 (D-Phe-Pip-Arg-pNA, Km=2.54 xcexcM).
Human FXIa assay (Enzyme Research Labs) was measured as for FXa except that the reaction was started with 100 xcexcM S-2366 (L-pyroGlu-Pro-Arg-pNA, Km=86 xcexcM).
Bovine and human pancreatic trypsin (Sigma) were assayed in 2 mM CaCl2, 50 mM Tris/Cl (pH 8.0) and the reaction was started with 100 xcexcM Chromozym-TRY (Carboxybenzoxy-Val-Gly-Arg-pNA, Km=46 xcexcM).
Tryptase inhibition activity was measured using either isolated human skin tryptase or recombinant human tryptase prepared from the human recombinant beta-protryptase expressed by baculovirus in insect cells. The expressed beta-protryptase to was purified using sequential immobilized heparin affinity resin followed by an immunoaffinity column using an anti-tryptase monoclonal antibody. The protryptase was activated by auto-catalytic removal of the N-terminal in the presence of dextran sulfate followed by dipeptidyl peptidase I (DPPI) removal of the two N-terminal amino acids to give the mature active enzyme (Sakai et al, J. Clin. Invest., Vol. 97 (1996), at pp. 988-995). Essentially equivalent results were obtained using isolated native enzyme or the activated expressed enzyme. The tryptase enzyme was maintained in 2M sodium chloride, 10 nM 4-morpholine-propanesulfonic acid, pH 6.8. The assay procedure employed a 96 well microplate. To each well of the microplate (Nunc MaxiSorp), 250 xcexcl of assay buffer [containing low molecular weight heparin and tris (hydroxymethyl)aminomethane] was added followed by 2.0 xcexcl of the test compound in dimethylsulfoxide. The substrate (10 xcexcl) was then added to each well to give a final concentration of 100 xcexcM benzyloxycarbonyl-glycine-proline-arginine-p-nitroaniline (CBz-Gly-Pro-Arg-pNA). The microplate was then shaken on a platform vortex mixer at a setting of 800 (Sarstedt TPM-2). After a total of three minutes incubation, 10 xcexcl of the working stock solution of tryptase was added to each well. The microplate was vortexed again for one minute and then incubated without shaking at RT for an additional 2 minutes. After this time the microplate was read on a microplate reader (Molecular Devices UV max) in the kinetic mode (405 nm wavelength) over twenty minutes at RT. To determine the compound concentration that inhibited half of the enzyme activity (IC50), the fraction of control activity (FCA) was plotted as a function of the inhibitor concentration and curve to fit FCA/(1[I]/IC50). The IC50 for each compound was determined 2-4 times and the obtained values were averaged.
Applying the above-described assays, the inventive compounds demonstrated activity as inhibitors of Factors VIIa, IXa, Xa, XIa, IXa, tryptase and/or urokinase.